Characterization of intracellular free Ca2+ movements in neural progenitor cells derived from ES cells transfected with MASH 1 transcription factor gene
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概要
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Intracellular free Ca2+ movements in neural progenitor cells obtained by transfection of embryonic stem(ES) cells with MASH1 transcription factor gene were studied. The MASH1 transfected cells, majority of which were Islet1 positive, have been shown to improve motor functions of hemiplegic mice when transplanted into the injured brain. In this study RT-PCR was conducted to detect Ca2+ channel mRNAs. We found that mRNAs of L-type, N-type and T-type Ca2+ channel mRNAs were expressed in the MASH1 transfected cells. Next, a Ca2+-sensitive fluorescence probe, fluo-3, and a scanning confocal laser microscope were used to detect any changes of intracellular Ca2+ concentration. Enhanced fluo-3 fluorescence was observed when the cells were stimulated with increased extracellular potassium, a depolarization signal. Depletion of extracellular Ca2+ abrogated the increase of fluorescence intensity upon depolarization. A Ca2+ channel blocker, lead, inhibited the increase of fluorescence intensity upon depolarization. A L-type channel inhibitor, nifedipine, but not a N-type channel inhibitor, omega-conotoxin GVIA, reduced the increase of fluorescence intensity upon depolarization. Thapsigargin, which depletes intracellular Ca2+ stores, did not attenuate the depolarization-induced signals. Potassium channel inhibitors, tetraethylammonium and 4-aminopyridine inhibited the fluorescence signals. These results indicate that depolarization-induced intracellular Ca2+ increase is mainly due to inward flow (influx) of Ca2+ via Ca2+ channels and marginally due to the release (mobilization) of Ca2+ from intracellular stores in the MASH1 transfected neural progenitor cells. The calcium signals may regulate various aspects of neural cell function such as authentic cellular activity including neurotransmitter secretion and cell differentiation. Our data add the possibility that calcium signals may play roles even in the ectopically transplanted MASH1-transfected neural progenitor cells reconstituting neural network to improve motor functions of hemiplegic mice.
- 2005-09-25
著者
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Suzuki Noboru
Department of Advanced Interdisciplinary Sciences, Graduate School of Engineering, Utsunomiya Univer
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HASHIMOTO Takuo
Department of Neurosurgery, St. Marianna University School of Medicine
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榊原 学
東海大学開発工学部生物工学科
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WATANABE Kenji
Department of Kampo Medicine, Keio University, School of Medicine
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Ueda Yuji
Department of Earth and Planetary Science, Graduate School of Science, University of Tokyo
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Suzuki Noboru
St. Marianna Univ. School Of Medicine Kawasaki Jpn
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Suzuki Noboru
Department Of Immunology And Medicine St. Marianna University School Of Medicine
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S. Kurokawa
Department Of Immunology And Medicine St. Marianna University School Of Medicine
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榊原 学
東海大 開発工
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Hashimoto Takuo
Department Of Neurosurgery St. Marianna University School Of Medicine
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Hashimoto Takuo
Department Of Neurological Surgery Tokyo Women's Medical College
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Ueda Yuji
Department Of Earth And Planetary Science University Of Tokyo
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YOSHIKAWA Hideshi
Department of Parasitology and Immunology, Yamanashi Medical University
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IDE Michiko
Department of Immunology and Medicine, St. Marianna University School of Medicine
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SAKAIBARA Manabu
Laboratory of Neurobiological Engineering, Department of Biological Science and Technology, School o
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Ide Michiko
Department Of Immunology And Medicine St. Marianna University School Of Medicine
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Ueda Yuji
Department Of Immunology And Medicine St. Marianna University School Of Medicine
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Watanabe Kenji
Department Of Gastroenterology Osaka City University Graduate School Of Medicine
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Yoshikawa Hideshi
Department Of Immunology And Medicine St. Marianna University School Of Medicine
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Suzuki Noboru
Department Of Advanced Interdisciplinary Sciences Graduate School Of Engineering Utsunomiya Universi
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Watanabe Kenji
Department Of Anatomy Fukui Medical School
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Watanabe Kenji
Department Of Immunology And Medicine St. Marianna University School Of Medicine
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Watanabe Kenji
Department Of Agricultural Chemistry College Of Agriculture And Veterinary Medicine Nihon University
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